针刺抗氧化效应的TRx氧化还原调控机制研究

项目名称： 针刺抗氧化效应的TRx氧化还原调控机制研究

项目编号： No.30801483

项目类型： 青年科学基金项目

立项/批准年度： 2009

项目学科： 金属学与金属工艺

项目作者： 刘存志

作者单位： 天津中医药大学

项目金额： 20万元

中文摘要： 本课题以自体栓子注入法复制血管性痴呆大鼠模型为研究对象，从TRx氧化还原调控通路探讨针刺改善脑功能的抗氧化机制。采用Morris水迷宫和小动物PET技术评价动物的学习记忆能力和脑代谢水平；分光光度法检测海马NADPH/NADP+和GSH/GSSG比值；采用实时定量RT-PCR、Western Blotting技术及分光光度法分别检测海马硫氧还蛋白TRx1的表达水平和硫氧还蛋白还原酶TRxR的活性变化；免疫组化及Western Blotting方法分析海马Ref-1、NF-κ、Iκ#21644;IKK的蛋白表达水平。研究发现，针刺可改善血管性痴呆大鼠空间学习记忆能力以及相关脑功能区的葡萄糖代谢率，通过提高海马CA1区、CA3区和齿状回Ref-1的蛋白表达水平，提高海马CA1区NF-κ#65293;IkB表达水平，发挥抗氧化和神经保护作用，这为针刺抗氧化作用机理的研究提供了实验依据。但针刺对Ref-1相关氧化还原信号通路的具体调控效应仍不清楚，将是课题组下一步研究的重点。

中文关键词： 针刺；海马；氧化还原；氧化还原因子-1；血管性痴呆

英文摘要： The aim of this project is to explore the antioxidative mechanism of acupuncture improving brain function through redox control pathway of thioredoxin (TRx). The vascular dementia model is made by injecting homogeneous emboli into the internal carotid artery in rats. The learning and memory performance and cerebral glucose metabolism level in different brain function regions are measured by Morris water maze test and small animal positron emission tomography (PET) technology, the ratios of NADPH/NADP+ and glutathione (GSH)/GSSG in hippocampus by spectrophotometric method, the expression level of TRx1 and activity change of thioredoxin reductase (TRxR) in hippocampus by real-time reverse transcription-polymerase chain reaction (RT-PCR),western blotting analysis and spectrophotometric method, and the protein expression level of redox factor 1 (Ref-1), NF-κIκ and IkB kinase (IKK) in hippocampus by immunohistochemistry and western blotting technologies. Our researches have demonstrated that acupuncture could alleviate the space learning and memory impairment induced by vascular dementia in rats, and improve the glucose metabolism rate in related brain regions. Acupuncture made an antioxidative and neuroprotective effect by raising the Ref-1 expression level in CA1、CA3and DG and elevating the level of NF-κ#65293;IkB expression in CA1. This provided scientific support for further study of acupuncture antioxidative effect.However, it is not yet clear about the specific regulation effect of acupuncture treatment on Ref-1 related redox signal pathway.

英文关键词： acupuncture; hippocampus; redox; Ref-1; vascular dementia